ALTERATIONS EN mRNA EXPRESSION OF APOPTOSIS-RELATED GENES AFTER TREATMENT OF SKOV-3 CELLS WITH CARDAMONIN

The aim of this study is to investigate the effect of cardamonin on mRNA expression of apoptosisrelated genes and proliferation after treatment of ovarian epithelial cells, SKOV-3. Real time cellular assay and acridine orange/propidium iodide staining were used to determine cell proliferation rate and viability in the presence and absence of cardamonin, respectively. The apoptosis and necrosis were quantified by annexin V/ propidium iodide uptake. In addition, the mRNA expression levels in cells containing 25-75 μM cardamonin were determined using RT-PCR. The results indicated that the pre-treatment of cells with 25‒150 μM cardamonin attenuated the cell proliferation rate and cell viability significantly in a dose-dependent manner. The early apoptotic cell number was increased significantly (p≤0, 01) by treatment of 50 μM cardamonin as compared the untreated cells after 72 h incubation. The analyses of mRNA expressions showed an increase in caspase-3 and Bax apoptotic genes after 25 μM cardamonin treatment versus untreated cells. However, the mRNA expression levels of Bcl-2, Bcl-xL, NF-κB and cyclin D1 were attenuated by cardamonin in cells. Taken together, cardamonin supressed the cells proliferation in a dose dependent manner. These effects could be explained by down regulation of mRNA expression of NF-κB and cyclin D1. In addition, cardamonin stimulated apoptosis by down regulation of anti-apoptotic genes and upregulation of caspase-3 and Bax in SKOV-3 cell line.